Characterization of a sperm-specific nuclear autoantigenic protein. I. Complete sequence and homology with the Xenopus protein, N1/N2.
In our studies on specific sperm proteins that function in fertilization, an autoantigenic, postacrosomal sperm protein has been found to originate in the testis as a nuclear-associated protein. This nuclear autoantigenic sperm protein (NASP) contains a C-terminal nuclear translocation signal and has structural similarities to the lamins and other nuclear proteins; and its 2.5 kb mRNA is apparently tissue-, but not species-, specific. DNA clones from a rabbit testis cDNA library and a rabbit genomic library were sequenced in order to characterize NASP. The polyadenylated mRNA has 39 bases of 5' untranslated sequence, an open reading frame of 2043 bases encoding 680 amino acids, and a 104 base 3' untranslated region (2,186). The encoded polypeptide has a calculated molecular weight of 73,533 and a pI = 4.06, containing 25% acidic residues. One clone (R1.2) expressing the C-terminal 446 amino acids was used to express a fusion protein. The expressed R1.2/beta-galactosidase fusion protein was found to be autoantigenic. Secondary structure predictions for NASP showed that 69% of the molecule had a high probability of forming alpha-helices and that several alpha-helical regions had a characteristic repeating heptad pattern that in the intermediate filaments and nuclear lamins is involved in coiled-coil interactions with other molecules. In addition to the nuclear translocation signal common to many nuclear proteins, NASP also showed homology with the Xenopus histone-binding protein, N1/N2.
Welch
J E
JE
Department of Cell Biology and Anatomy, University of North Carolina, Chapel Hill 27599.
Zimmerman
L J
LJ
Joseph
D R
DR
O'Rand
M G
MG
D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics; D018797:Cell Cycle Proteins; D002467:Cell Nucleus / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid; D013094:Spermatozoa / Q000737:chemistry* / Q000648:ultrastructure; D013737:Testis / Q000737:chemistry; D014981:Xenopus
D001324:Autoantigens; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C051899:N1-N2 protein, Xenopus; D009687:Nuclear Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C067535:Nasp protein, mouse; D004247:DNA